Immunocytochemical Analysis

Johnson, Margaret A.

doi:10.1385/1-59259-138-8:339

Cited by 2 publications

(2 citation statements)

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“…Optimised immunohistochemical and multiplex western blot protocols were used for the demonstration and resolution of muscular dystrophy associated proteins, as previously described. 22 23 Primary antibodies used for immunohistochemistry were directed against: β-spectrin (RBC2/3D5), β-dystroglycan (43DAG/8D5), C terminus dystrophin (Dy8/6C5) and N terminus dystrophin (Dy10/12B2), α-sarcoglycan (Ad1/20A6), β-sarcoglycan (1/5B1), γ-sarcoglycan (35DAG/21B5), δ-sarcoglycan (3/12C1) (all from Novocastra, Newcastle, UK) and caveolin-3 (Transduction Laboratories, Oxford, UK). Antibodies used for western blotting were: anti-dysferlin (NCL-hamlet exon 53), anti-dystrophin (Dy8/6C5 C-terminus and Dy4/6D3 rod domain), anti-α-sarcoglycan, anti-β-dystroglycan, anti-γ-sarcoglycan, anti-calpain 3 (NCL-CALP-2C4 and NCL-CALP-12A2) and anti-caveolin 3 (Transduction Laboratories).…”

Section: Methodsmentioning

confidence: 99%

New aspects on patients affected by dysferlin deficient muscular dystrophy

Klinge

Aboumousa

Eagle

et al. 2009

Journal of Neurology, Neurosurgery & Psychiatry

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Mutations in the dysferlin gene lead to limb girdle muscular dystrophy 2B, Miyoshi myopathy and distal anterior compartment myopathy. A cohort of 36 patients affected by dysferlinopathy is described, in the first UK study of clinical, genetic, pathological and biochemical data. The diagnosis was established by reduction of dysferlin in the muscle biopsy and subsequent mutational analysis of the dysferlin gene. Seventeen mutations were novel; the majority of mutations were small deletions/insertions, and no mutational hotspots were identified. Sixty-one per cent of patients (22 patients) initially presented with limb girdle muscular dystrophy 2B, 31% (11 patients) with a Miyoshi phenotype, one patient with proximodistal mode of onset, one patient with muscle stiffness after exercise and one patient as a symptomatic carrier. A wider range of age of onset was noted than previously reported, with 25% of patients having first symptoms before the age of 13 years. Independent of the initial mode of presentation, in our cohort of patients the gastrocnemius muscle was the most severely affected muscle leading to an inability to stand on tiptoes, and lower limbs were affected more severely than upper limbs. As previous anecdotal evidence on patients affected by dysferlinopathy suggests good muscle prowess before onset of symptoms, we also investigated pre-symptomatic fitness levels of the patients. Fifty-three per cent of the patients were very active and sporty before the onset of symptoms which makes the clinical course of dysferlinopathy unusual within the different forms of muscular dystrophy and provides a challenge to understanding the underlying pathomechanisms in this disease.

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Section: Methodsmentioning

confidence: 99%

New aspects on patients affected by dysferlin deficient muscular dystrophy

Klinge

Aboumousa

Eagle

et al. 2009

Journal of Neurology, Neurosurgery & Psychiatry

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“…Immunolabelling of tissue sections and blots was performed as described previously [28,29]. Muscle biopsy samples are examined using the antibodies listed in Table 1.…”

Section: Immunolabellingmentioning

confidence: 99%